Aberration correction for real-time measurements in adaptive confocal microscopy

• Applicants: Professor Dr.-Ing. Jürgen W. Czarske; Dr. Stefan Scholz; Professorin Dr.-Ing. Ulrike Wallrabe
• Subject Area: Measurement Systems ; Microsystems
• Term: from 2015 to 2024
• Website: Homepage
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 271021903

• Final Report Year: 2024

Final Report Abstract

In the present research project, a fully adaptive confocal microscope was realized, which relies exclusively on adaptive optical elements, completely eliminating the need for mechanically moving parts. The adaptive elements simultaneously enable a 3D scan and aberration correction. Initially, novel adaptive lenses with spherical aberration correction were investigated and employed for axial scanning. Through in vivo fluorescence measurements in zebrafish embryos, it was demonstrated that such lenses allow simultaneous tuning of focal length and correction of both symmetric (spherical) and asymmetric (astigmatism and coma) sample-induced aberrations. Building upon these results, diffraction-limited scanning was extended to three dimensions. For this purpose, a biaxial adaptive prism was developed, enabling lateral scanning in transmission. Combined with an adaptive lens, a fully adaptive microscope was realized. To achieve homogeneous focus quality across a large three-dimensional scanning range, a telecentric F- Theta lens was specifically designed and implemented. Fluorescence measurements on zebrafish embryos successfully demonstrated the feasibility of fully adaptive, high-resolution three-dimensional scanning. In fluorescence measurements, it is crucial to address chromatic aberrations due to the use of at least two wavelengths. Consequently, an adaptive lens with additional chromatic aberration correction was implemented, and its functionality was successfully demonstrated. One of the major challenges in the application of adaptive elements lies in their precise control. This was predominantly achieved through a closed-loop system, where the behavior of adaptive elements was measured using digital holography, and voltages were regulated accordingly. Towards the end of the project, successful adoption of machine learning methods, as an alternative to classical control techniques, facilitated precise control of the adaptive achromat. In summary, this project successfully showcased that adaptive optical elements can significantly contribute to three-dimensional high-resolution fluorescence microscopy. Their capability for flexible scanning and simultaneous correction of sample-induced aberrations represents a substantial advancement in imaging. The integration of machine learning also opens promising avenues for the development of intelligent microscopes, establishing a crucial foundation for future technologies in this field.

Publications

• High-contrast 3D image acquisition using HiLo microscopy with an electrically tunable lens. SPIE Proceedings, 9890, 98900A.
  Philipp, Katrin; Smolarski, André; Fischer, Andreas; Koukourakis, Nektarios; Stürmer, Moritz; Wallrabe, Ulricke & Czarske, Jürgen
  
• Topological in-plane polarized piezo actuation for compact adaptive lenses with aspherical correction, ACTUATOR 2016, 15th International Conference on New Actuators, Bremen, Germany, pp. 228-231 (2016)
  F. Lemke; M. Stürmer; U. Wallrabe & M.C. Wapler
  
• Volumetric HiLo microscopy employing an electrically tunable lens. Optics Express, 24(13), 15029.
  Philipp, Katrin; Smolarski, André; Koukourakis, Nektarios; Fischer, Andreas; Stürmer, Moritz; Wallrabe, Ulrike & Czarske, Jürgen W.
  
• Adaptive lenses for axial scanning in HiLo microscopy. Optics in the Life Sciences Congress, BoTu1A.2.
  Koukourakis, Nektarios; Philipp, Katrin; Lemke, Florian; Stürmer, Moritz; Wapler, Matthias; Wallrabe, Ulrike & Czarske, Jürgen
  
• Spherical aberration correction of adaptive lenses. SPIE Proceedings, 10073, 1007303.
  Philipp, Katrin; Lemke, Florian; Wapler, Matthias C.; Wallrabe, Ulrike; Koukourakis, Nektarios & Czarske, Jürgen W.
  
• Structured illumination 3D microscopy using adaptive lenses and multimode fibers. SPIE Proceedings, 10335, 1033519.
  Czarske, Jürgen; Philipp, Katrin & Koukourakis, Nektarios
  
• “Adaptive hybrid illumination microscopy employing electrically tunable lenses with aberration correction”, 24th Congress of the International Commission for Optics in Tokyo, Japan, 21.08. - 25.08.2017
  K. Philipp; N Koukourakis; F. Lemke; M. Wapler; U. Wallrabe & Jürgen W. Czarske
  
• “Adaptive hybrid illumination microscopy for Zebrafish screening”, Speckle VII 2018, 10. - 12.9.2018, Janow Podlaski (PL)
  N. Koukourakis; K. Philipp; M. Stürmer; U. Wallrabe & J.W. Czarske
  
• “Characterization of adaptive lenses using partitioned aperture wavefront imaging“, European Optical Society Biennial Meeting (EOSAM) 2018, 8. – 12.10.2018, Delft, Netherlands
  W. Wang; K. Philipp; N. Koukourakis & J. Czarske
  
• Adaptive hybrid illumination microscopy for Zebrafish screening. Biophotonics Congress: Optics in the Life Sciences Congress 2019 (BODA,BRAIN,NTM,OMA,OMP), NT3C.2.
  Koukourakis, Nektarios & Czarske, Jürgen
  
• Axial scanning and spherical aberration correction in confocal microscopy employing an adaptive lens (Conference Presentation). Adaptive Optics and Wavefront Control for Biological Systems V, 13.
  Czarske, Jürgen W.; Lemke, Florian; Wapler, Matthias C.; Wallrabe, Ulricke; Koukourakis, Nektarios & Philipp, Katrin
  
• Axial scanning employing tunable lenses: Fourier optics based system design. OSA Continuum, 2(4), 1318.
  Philipp, Katrin & Czarske, Jürgen
  
• Diffraction-limited axial scanning in thick biological tissue with an aberration-correcting adaptive lens. Scientific Reports, 9(1).
  Philipp, Katrin; Lemke, Florian; Scholz, Stefan; Wallrabe, Ulrike; Wapler, Matthias C.; Koukourakis, Nektarios & Czarske, Jürgen W.
  
• Miniaturized piezo-actuated adaptive prism for biaxial optical scanning. 2019 International Conference on Optical MEMS and Nanophotonics (OMN), 222-223.
  Lemke, Florian; Weber, Pascal M.; Wallrabe, Ulrike & Wapler, Matthias C.
  
• Multiphysics simulation of the aspherical deformation of piezo-glass membrane lenses including hysteresis, fabrication and nonlinear effects. Smart Materials and Structures, 28(5), 055024.
  Lemke, Florian; Frey, Yasmina; Bruno, Binal P.; Philipp, Katrin; Koukourakis, Nektarios; Czarske, Jürgen; Wallrabe, Ulrike & Wapler, Matthias C.
  
• Quantitative phase imaging for in-situ monitoring of adaptive lenses. Digital Holography and Three-Dimensional Imaging 2019, M4B.5.
  Koukourakis, Nektarios; Wang, Wenjie; Philipp, Katrin & Czarske, Jürgen
  
• Ultra-Fast and Compact Varifocal Lens. 2019 IEEE 32nd International Conference on Micro Electro Mechanical Systems (MEMS), 938-941.
  Wapler, Matthias C & Wallrabe, Ulrike
  
• Piezo-actuated adaptive elements in scanning microscopy (Conference Presentation). Unconventional Optical Imaging II, 50.
  Wang, Wenjie; Koukourakis, Nektarios & Czarske, Jürgen W.
  
• Piezo-actuated adaptive prisms for continuously adjustable bi-axial scanning. Smart Materials and Structures, 29(9), 095004.
  Lemke, Florian; Weber, Pascal M.; Philipp, Katrin; Czarske, Jürgen W.; Koukourakis, Nektarios; Wallrabe, Ulrike & Wapler, Matthias C.
  
• Real-time monitoring of adaptive lenses with high tuning range and multiple degrees of freedom. Optics Letters, 45(2), 272.
  Wang, Wenjie; Philipp, Katrin; Koukourakis, Nektarios & Czarske, Jürgen W.
  
• 3D-scanning microscopy with adaptive lenses and prisms for zebrafish studies. Journal of Optical Microsystems, 1(02).
  Wang, Wenjie; Lemke, Florian; Wapler, Matthias C.; Wallrabe, Ulrike; Czarske, Jürgen W. & Koukourakis, Nektarios
  
• A One-Inch Aperture Piezoelectric Tunable Lens with Small Footprint. 2021 21st International Conference on Solid-State Sensors, Actuators and Microsystems (Transducers), 427-430.
  Gowda, Hitesh G. B.; Graf, Tobias & Wallrabe, Ulrike
  
• Einstellbare Prismen mit Glas-Luft Interface: Ein Vergleich verschiedener Aktuierungsmethoden, Mikrosystemtechnikkongress 2021, Ludwigsburg, Germany, pp. 306–309 (2021)
  P. M. Weber; F. Lemke & M. C. Wapler
  
• A 3D scanner: low footprint piezoelectric tunable optical scanner. MOEMS and Miniaturized Systems XXI, 15.
  Gowda, Hitesh G. B.; Gräf, Tobias & Wallrabe, Ulrike
  
• Assignment of Focus Position with Convolutional Neural Networks in Adaptive Lens Based Axial Scanning for Confocal Microscopy. Applied Sciences, 12(2), 661.
  Schmidt, Katharina; Koukourakis, Nektarios & Czarske, Jürgen
  
• Tunable doublets: piezoelectric glass membrane lenses with an achromatic and spherical aberration control. Optics Express, 30(26), 46528.
  Gowda, Hitesh G. B.; Wapler, Matthias C. & Wallrabe, Ulrike
  
• Chromatic aberration correction employing reinforcement learning. Optics Express, 31(10), 16133.
  Schmidt, Katharina; Guo, Ning; Wang, Wenjie; Czarske, Juergen & Koukourakis, Nektarios
  
• Chromatic aberration correction using reinforcement learning. Emerging Topics in Artificial Intelligence (ETAI) 2023, 35.
  Schmidt, Katharina; Guo, Ning; Wang, Wenjie; Czarske, Juergen W. & Koukourakis, Nektarios
  
• Fully refractive telecentric f-theta microscope based on adaptive elements for 3D raster scanning of biological tissues. Optics Express, 31(18), 29703.
  Wang, Wenjie; Schmidt, Katharina; Wapler, Matthias C.; Wallrabe, Ulrike; Czarske, Jürgen W. & Koukourakis, Nektarios
  
• Higher order wavefront correction and axial scanning in a single fast and compact piezo-driven adaptive lens. Optics Express, 31(14), 23393.
  Gowda, Hitesh G. B.; Wallrabe, Ulrike & Wapler, Matthias C.
  
• Piezo-actuated higher order wavefront correction lens, TRANSDUCERS 2023, TRANSDUCERS 2023, Kyoto, Jp., pp. 1477-1479 (2023)
  H.G.B Gowda; M.C. Wapler & U. Wallrabe
  
• Reliability of tunable lenses: feedback sensors and the influence of temperature, orientation, and vibrations. Applied Optics, 62(12), 3072.
  Gowda, Hitesh G. B.; Bruno, Binal P.; Wapler, Matthias C. & Wallrabe, Ulrike